Phosphorylation and activation of PKCδ by Src is important for the scatter of MDCK cells upon HGF stimulation A. MDCK cells were serum-starved for 24 h and were then treated with (+) or without (−) the Src inhibitor dasatinib at 100 nM for 1 h before they were stimulated with HGF (20 ng/ml) for 15 min. Endogenous PKCδ was immunoprecipitated using an anti-PKCδ antibody, and the immunocomplexes were analyzed by immunoblotting with antibodies to PKCδ or PKCδ pY311. To measure the PKCδ activity, the immuoncomplexes were subjected to an in vitro kinase assay in the presence of [γ- 32 P]ATP and myelin basic protein (MBP) as the substrate. The 32 P-incorporated MBP were fractionated by SDS-polyacrylamide gel electrophoresis and visualized by autoradiography. The radioisotope activity was quantified using a phosphoimager system. The data are expressed as fold relative to the level of the control. Values (mean ± SD) are from three experiments. *, P
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